Inclusion of IL-3 during retrovirally-mediated transduction on stromal support does not increase the extent of gene transfer into long-term engrafting human hematopoietic progenitors

Mo A. Dao, Jan Nolta

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Interleukin 3 (IL-3) supports the survival of multilineage hematopoietic progenitors, and increases the extent of retrovirally mediated gene transfer into colony-forming cells. However, effects from the supraphysiological levels used in ex-vivo expansion and gene-therapy protocols on subsequent differentiation of human progenitors have not been well defined. In the current studies, the extents of retrovirally mediated transduction and lineage development from CD34+ cells cultured ex vivo in the presence or absence of IL-3 were compared. All transductions were performed in the presence of an irradiated stromal support layer, IL-6 and SCF, with and without inclusion of 10 ng/ml IL-3. Following transduction, colony-forming (CFU) assays were done, and the remaining cells were transplanted into cohorts of sibling beige/nude/xid (bnx) mice. Marrow from the mice was harvested 9-10 months post transplantation. The average extent of human CD45+ cell engraftment in the bone marrow and the human hematopoietic lineages recovered from the mice in the +IL-3 and -IL-3 groups did not vary significantly. No deleterious effects on the extent of engraftment, lineage generation, or survival of clonogenic progenitors was observed with inclusion of IL-3 in the transductions performed on stromal support. The percentage of G418-resistant human progenitors recovered from mice was equivalent. The extent of marking by the neo gene in the marrow of the mice was equal in both groups, and inverse PCR revealed that primitive cells transduced in the absence of IL-3 had generated progeny with slightly better clonal diversity than progenitors transduced in the presence of IL-3. These data show that, while transduction of colony-forming progenitors may not always be apparent, primitive human hematopoietic cells can be transduced to significant levels in the absence of IL-3.

Original languageEnglish (US)
Pages (from-to)81-89
Number of pages9
JournalCytokines, Cellular and Molecular Therapy
Volume3
Issue number2
StatePublished - Jun 1997
Externally publishedYes

Fingerprint

Interleukin-3
Genes
Bone Marrow
Nude Mice
Genetic Therapy
Cultured Cells
Interleukin-6
Transplantation
Polymerase Chain Reaction
Survival

Keywords

  • Gene therapy
  • Immune-deficient mice
  • Interleukin-3
  • Retroviral vector
  • Transduction

ASJC Scopus subject areas

  • Pharmacology
  • Immunology and Allergy
  • Immunology

Cite this

@article{c080044e782b41f1a95c55867b8d89a3,
title = "Inclusion of IL-3 during retrovirally-mediated transduction on stromal support does not increase the extent of gene transfer into long-term engrafting human hematopoietic progenitors",
abstract = "Interleukin 3 (IL-3) supports the survival of multilineage hematopoietic progenitors, and increases the extent of retrovirally mediated gene transfer into colony-forming cells. However, effects from the supraphysiological levels used in ex-vivo expansion and gene-therapy protocols on subsequent differentiation of human progenitors have not been well defined. In the current studies, the extents of retrovirally mediated transduction and lineage development from CD34+ cells cultured ex vivo in the presence or absence of IL-3 were compared. All transductions were performed in the presence of an irradiated stromal support layer, IL-6 and SCF, with and without inclusion of 10 ng/ml IL-3. Following transduction, colony-forming (CFU) assays were done, and the remaining cells were transplanted into cohorts of sibling beige/nude/xid (bnx) mice. Marrow from the mice was harvested 9-10 months post transplantation. The average extent of human CD45+ cell engraftment in the bone marrow and the human hematopoietic lineages recovered from the mice in the +IL-3 and -IL-3 groups did not vary significantly. No deleterious effects on the extent of engraftment, lineage generation, or survival of clonogenic progenitors was observed with inclusion of IL-3 in the transductions performed on stromal support. The percentage of G418-resistant human progenitors recovered from mice was equivalent. The extent of marking by the neo gene in the marrow of the mice was equal in both groups, and inverse PCR revealed that primitive cells transduced in the absence of IL-3 had generated progeny with slightly better clonal diversity than progenitors transduced in the presence of IL-3. These data show that, while transduction of colony-forming progenitors may not always be apparent, primitive human hematopoietic cells can be transduced to significant levels in the absence of IL-3.",
keywords = "Gene therapy, Immune-deficient mice, Interleukin-3, Retroviral vector, Transduction",
author = "Dao, {Mo A.} and Jan Nolta",
year = "1997",
month = "6",
language = "English (US)",
volume = "3",
pages = "81--89",
journal = "Cytokines, Cellular and Molecular Therapy",
issn = "1368-4736",
publisher = "Taylor and Francis Ltd.",
number = "2",

}

TY - JOUR

T1 - Inclusion of IL-3 during retrovirally-mediated transduction on stromal support does not increase the extent of gene transfer into long-term engrafting human hematopoietic progenitors

AU - Dao, Mo A.

AU - Nolta, Jan

PY - 1997/6

Y1 - 1997/6

N2 - Interleukin 3 (IL-3) supports the survival of multilineage hematopoietic progenitors, and increases the extent of retrovirally mediated gene transfer into colony-forming cells. However, effects from the supraphysiological levels used in ex-vivo expansion and gene-therapy protocols on subsequent differentiation of human progenitors have not been well defined. In the current studies, the extents of retrovirally mediated transduction and lineage development from CD34+ cells cultured ex vivo in the presence or absence of IL-3 were compared. All transductions were performed in the presence of an irradiated stromal support layer, IL-6 and SCF, with and without inclusion of 10 ng/ml IL-3. Following transduction, colony-forming (CFU) assays were done, and the remaining cells were transplanted into cohorts of sibling beige/nude/xid (bnx) mice. Marrow from the mice was harvested 9-10 months post transplantation. The average extent of human CD45+ cell engraftment in the bone marrow and the human hematopoietic lineages recovered from the mice in the +IL-3 and -IL-3 groups did not vary significantly. No deleterious effects on the extent of engraftment, lineage generation, or survival of clonogenic progenitors was observed with inclusion of IL-3 in the transductions performed on stromal support. The percentage of G418-resistant human progenitors recovered from mice was equivalent. The extent of marking by the neo gene in the marrow of the mice was equal in both groups, and inverse PCR revealed that primitive cells transduced in the absence of IL-3 had generated progeny with slightly better clonal diversity than progenitors transduced in the presence of IL-3. These data show that, while transduction of colony-forming progenitors may not always be apparent, primitive human hematopoietic cells can be transduced to significant levels in the absence of IL-3.

AB - Interleukin 3 (IL-3) supports the survival of multilineage hematopoietic progenitors, and increases the extent of retrovirally mediated gene transfer into colony-forming cells. However, effects from the supraphysiological levels used in ex-vivo expansion and gene-therapy protocols on subsequent differentiation of human progenitors have not been well defined. In the current studies, the extents of retrovirally mediated transduction and lineage development from CD34+ cells cultured ex vivo in the presence or absence of IL-3 were compared. All transductions were performed in the presence of an irradiated stromal support layer, IL-6 and SCF, with and without inclusion of 10 ng/ml IL-3. Following transduction, colony-forming (CFU) assays were done, and the remaining cells were transplanted into cohorts of sibling beige/nude/xid (bnx) mice. Marrow from the mice was harvested 9-10 months post transplantation. The average extent of human CD45+ cell engraftment in the bone marrow and the human hematopoietic lineages recovered from the mice in the +IL-3 and -IL-3 groups did not vary significantly. No deleterious effects on the extent of engraftment, lineage generation, or survival of clonogenic progenitors was observed with inclusion of IL-3 in the transductions performed on stromal support. The percentage of G418-resistant human progenitors recovered from mice was equivalent. The extent of marking by the neo gene in the marrow of the mice was equal in both groups, and inverse PCR revealed that primitive cells transduced in the absence of IL-3 had generated progeny with slightly better clonal diversity than progenitors transduced in the presence of IL-3. These data show that, while transduction of colony-forming progenitors may not always be apparent, primitive human hematopoietic cells can be transduced to significant levels in the absence of IL-3.

KW - Gene therapy

KW - Immune-deficient mice

KW - Interleukin-3

KW - Retroviral vector

KW - Transduction

UR - http://www.scopus.com/inward/record.url?scp=0030827301&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030827301&partnerID=8YFLogxK

M3 - Article

C2 - 9287247

AN - SCOPUS:0030827301

VL - 3

SP - 81

EP - 89

JO - Cytokines, Cellular and Molecular Therapy

JF - Cytokines, Cellular and Molecular Therapy

SN - 1368-4736

IS - 2

ER -